Nitric oxide (NO) derived from endothelial NO synthase (eNOS) is an integral mediator of vascular control during muscle contractions. However, it is not known whether neuronal NOS (nNOS)-derived NO regulates tissue hyperaemia in healthy subjects, particularly during exercise. We tested the hypothesis that selective nNOS inhibition would reduce blood flow and vascular conductance (VC) in rat hindlimb locomotor muscle(s), kidneys and splanchnic organs at rest and during dynamic treadmill exercise (20 m min −1 , 10% grade). Nineteen male Sprague-Dawley rats (555 ± 23 g) were assigned to either rest (n = 9) or exercise (n = 10) groups. Blood flow and VC were determined via radiolabelled microspheres before and after the intra-arterial administration of the selective nNOS inhibitor S-methyl-l-thiocitrulline (SMTC, 2.1 ± 0.1 μmol kg −1 ). Total hindlimb muscle blood flow (control: 20 ± 2 ml min −1 100g −1 , SMTC: 12 ± 2 ml min −1 100g −1 , P < 0.05) and VC (control: 0.16 ± 0.02 ml min −1 100 g −1 mmHg −1 , SMTC: 0.09 ± 0.01 ml min −1 100 g −1 mmHg −1 , P < 0.05) were reduced substantially at rest. Moreover, the magnitude of the absolute reduction in blood flow and VC correlated (P < 0.05) with the proportion of oxidative muscle fibres found in the individual muscles or muscle parts of the hindlimb. During exercise, total hindlimb blood flow (control: 108 ± 7 ml min −1 100 g −1 , SMTC: 105 ± 8 ml min −1 100 g −1 ) and VC (control: 0.77 ± 0.06 ml min −1 100g −1 mmHg −1 ; SMTC: 0.70 ± 0.05 ml min −1 100g −1 mmHg −1 ) were not different (P > 0.05) between control and SMTC conditions. SMTC reduced (P < 0.05) blood flow and VC at rest and during exercise in the kidneys, adrenals and liver. These results enhance our understanding of the role of NO-mediated circulatory control by demonstrating that nNOS does not appear to subserve an obligatory role in the exercising muscle hyperaemic response in the rat.

Reactive oxygen species, such as hydrogen peroxide (H(2)O(2)), exert a critical regulatory role on skeletal muscle function. Whether acute increases in H(2)O(2) modulate muscle microvascular O(2) delivery-utilization (Qo(2)/Vo(2)) matching [i.e., microvascular partial pressure of O(2) (Pmv(O(2)))] at rest and following the onset of contractions is unknown. The hypothesis was tested that H(2)O(2) treatment (exogenous H(2)O(2)) would enhance Pmv(O(2)) and slow Pmv(O(2)) kinetics during contractions compared with control. Anesthetized, healthy young Sprague-Dawley rats had their spinotrapezius muscles either exposed for measurement of blood flow (and therefore QO(2)), VO(2), and Pmv(O(2)), or exteriorized for measurement of force production. Electrically stimulated twitch contractions (1 Hz, ~7 V, 2-ms pulse duration, 3 min) were evoked following acute superfusion with Krebs-Henseleit (control) and H(2)O(2) (100 μM). Relative to control, H(2)O(2) treatment elicited disproportionate increases in QO(2) and VO(2) that elevated Pmv(O(2)) at rest and throughout contractions and slowed overall Pmv(O(2)) kinetics (i.e., ~85% slower mean response time; P < 0.05). Accordingly, H(2)O(2) resulted in ~33% greater overall Pmv(O(2)), as assessed by the area under the Pmv(O(2)) curve (P < 0.05). Muscle force production was not altered with H(2)O(2) treatment (P > 0.05), evidencing reduced economy during contractions (~40% decrease in the force/VO(2) relationship; P < 0.05). These findings indicate that, although increasing the driving force for blood-myocyte O(2) flux (i.e., Pmv(O(2))), transient elevations in H(2)O(2) impair skeletal muscle function (i.e., reduced economy during contractions), which mechanistically may underlie, in part, the reduced exercise tolerance in conditions associated with oxidative stress.

Age-related increases in oxidative stress contribute to impaired skeletal muscle vascular control. However, recent evidence indicates that antioxidant treatment with tempol (4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl) attenuates flow-mediated vasodilation in isolated arterioles from the highly oxidative soleus muscle of aged rats. Whether antioxidant treatment with tempol evokes similar responses in vivo at rest and during exercise in senescent individuals and whether this effect varies based on muscle fiber type composition are unknown. We tested the hypothesis that redox modulation via acute systemic tempol administration decreases vascular conductance (VC) primarily in oxidative hindlimb locomotor muscles at rest and during submaximal whole body exercise (treadmill running at 20 m/min, 5% grade) in aged rats. Eighteen old (25-26 mo) male Fischer 344 x Brown Norway rats were assigned to either rest (n = 8) or exercise (n = 10) groups. Regional VC was determined via radiolabeled microspheres before and after intra-arterial administration of tempol (302 μmol/kg). Tempol decreased mean arterial pressure significantly by 9% at rest and 16% during exercise. At rest, similar VC in 26 out of 28 individual hindlimb muscles or muscle parts following tempol administration compared with control resulted in unchanged total hindlimb muscle VC (control: 0.18 ± 0.02; tempol: 0.17 ± 0.05 ml·min(-1)·100 g(-1)·mmHg(-1); P > 0.05). During exercise, all individual hindlimb muscles or muscle parts irrespective of fiber type composition exhibited either an increase or no change in VC with tempol (i.e., ↑11 and ↔17 muscles or muscle parts), such that total hindlimb VC increased by 25% (control: 0.93 ± 0.04; tempol: 1.15 ± 0.09 ml·min(-1)·100 g(-1)·mmHg(-1); P ≤ 0.05). These results demonstrate that acute systemic administration of the antioxidant tempol significantly impacts the control of regional vascular tone in vivo presumably via redox modulation and improves skeletal muscle vasodilation independently of fiber type composition during submaximal whole body exercise in aged rats.

Excess reactive oxygen species are implicated in the impaired peripheral vascular function evident during exercise in older individuals. We tested the hypothesis that an acute infusion of the antioxidant ascorbic acid (AA) in old rats would improve antioxidant capacity and reduce oxidative stress and, therefore, elevate hindlimb muscle blood flow at rest and during treadmill exercise in muscles containing principally type I and IIa muscle fibers. Total and individual hindlimb skeletal muscle blood flow was measured (radiolabeled microspheres) in old rats (26-28 months) at rest (n = 8) and during treadmill exercise (n = 8; 20 m·min⁻¹, 5% grade) before and after AA treatment (76 mg·(kg body mass)⁻¹ intra-arterial (i.a.) injection). AA elevated total antioxidant capacity (rest, ~37%; and exercise, 31%) and reduced oxidative stress (~26%, exercise only). AA reduced resting total hindlimb muscle blood flow (control, 25 ± 3; AA, 16 ± 2 mL·min⁻¹·(100 g)⁻¹; p < 0.05) and blood flow to 8 of 28 individual muscles with no fiber-type correlation (p > 0.05). During exercise there was no effect of AA on total hindlimb muscle blood flow (control, 154 ± 14; AA, 162 ± 13 mL·min⁻¹·(100 g)⁻¹; p > 0.05) or blood flow to any individual muscle. This disconnect between whole-body antioxidant status and skeletal muscle blood flow in old rats mandates consideration when pursuing antioxidant treatments experimentally or clinically in older populations.

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